Background: Acute myocardial infarction (AMI) remains a leading cause of global morbidity and mortality, with early diagnosis being crucial for effective treatment and improved patient outcomes. The search for reliable early biomarkers to supplement traditional cardiac markers has been ongoing, with heart-type fatty acid-binding protein (H-FABP) emerging as a potential candidate due to its rapid post-ischemic release into the bloodstream. Objective: This study aimed to evaluate the diagnostic accuracy of H-FABP in comparison to conventional cardiac troponin I for the early detection of AMI in patients presenting with acute chest pain. Methods: A descriptive cross-sectional study was conducted at the Chemical Pathology Department of Combined Military Hospital Lahore from December 21, 2022, to April 21, 2023. Eighty patients presenting with symptoms suggestive of AMI were enrolled. Serum concentrations of H-FABP and cardiac troponin I were measured using the sandwich ELISA method. Diagnostic performance was assessed in terms of sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and receiver operating characteristic (ROC) curves. Statistical analysis was performed using SPSS version 25. Results: H-FABP demonstrated a sensitivity of 96.2%, specificity of 60.0%, PPV of 91.3%, and NPV of 81.8%. In contrast, cardiac troponin I had a sensitivity of 81.3% and a specificity of 60.0%, with PPV and NPV values of 91.3% and 81.8%, respectively. The area under the ROC curve (AUC) for H-FABP was 0.640 (p=0.031), slightly higher than that for cardiac troponin I, which was 0.624 (p=0.057). Conclusion: H-FABP shows promise as a more sensitive early biomarker for AMI compared to cardiac troponin I, suggesting its potential utility in enhancing early diagnosis and improving patient care. However, its lower specificity highlights the need for its combined use with traditional markers for a more accurate diagnosis.